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Oxidative Medicine and Cellular Longevity
Volume 2017 (2017), Article ID 6412682, 14 pages
Research Article

Mitochondria-Targeted Antioxidants SkQ1 and MitoTEMPO Failed to Exert a Long-Term Beneficial Effect in Murine Polymicrobial Sepsis

1Ludwig Boltzmann Institute for Experimental and Clinical Traumatology in the Trauma Research Center of AUVA, Vienna, Austria
2Institute of Surgical Research, Faculty of Medicine, University of Szeged, Szeged, Hungary
3Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Jena, Germany
4A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Leninskie Gory, Moscow 119992, Russia
5Center for Sepsis Control and Care, Jena University Hospital, Jena, Germany

Correspondence should be addressed to Marcin F. Osuchowski

Received 30 March 2017; Revised 22 June 2017; Accepted 2 August 2017; Published 19 September 2017

Academic Editor: Moh H. Malek

Copyright © 2017 Pia Rademann et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Fig. 1. Score sheet for a custom-developed mouse clinical assessment scoring system. The well-being of mice is assessed based on six endpoints every 12 hours (beginning at 12h post-CLP); tree severity grades (i.e. 0, 1 or 2 points) are assigned. Euthanasia is indicated when: score ≥8 and/or by inability to trigger the startle reflex and/or BT<28°C (recorded in at least two sequential measurements). Supplementary Fig. 2. Comparison of body temperature (BT) profiles in CLP experiments performed in two different laboratories. BT profile was used as a surrogate of outcome to optimally standardize the magnitude of CLP severity between the surgery performed in Ludwig Boltzmann Institute (LBI; Vienna, Austria) and Institute of Surgical Research (ISR), University of Szeged (Hungary). Mice underwent three CLP runs at ISR and their BT profiles (at 6h, 12h and 24h) were compared to the BT profile (at 6 and 24h post-CLP) of the CLP mice enrolled in the survival study performed in LBI (solid line/dot). ISR-CLP #3 was designated as the best match and expanded to n⁼28 for further mitophagy analysis. LBI-CLP n⁼90-78; ISR-CLP#1 n⁼28-16; ISR-CLP#2 n⁼25-13; ISR-CLP#3 n⁼27-26. Data points shown as mean±SEM. Supplementary Fig. 3. Effect of SkQ1 treatment on the state 3 mitochondrial respiration in the rat and mouse liver homogenates. State 3 respiration was measured in mouse and rat liver homogenates subjected to SkQ1 in the range of concentrations from 0 to 108.5nM. n⁼4/each species. Data points shown as mean±SD. Dotted lines indicate either the single (5nM) or cumulative (25nM) SkQ1 dose administered to CLP mice in the main survival study (Experiment 1). The rat data serve as species comparison. Supplementary Fig. 4. Visualization of outcome for each individual CLP run. CLP was performed in six independent reiterations with 14-15 mice at each repetition (typically 5 mice/each group; the precise n indicated on each panel). Statistical assessment of outcome was performed on the combined data set (Fig. 2). Supplementary Fig. 5. Trajectory of white blood cells (WBC; A), lymphocyte (LYM; B) and neutrophil (NEU; C) counts for SkQ1, MitoTEMPO and placebo mice (control). For A-C: at BL n⁼50; at 6h control n⁼20, SkQ1 n⁼25, MitoTEMPO n⁼23; at 24h control n⁼23, SkQ1 n⁼23, MitoTEMPO n⁼22; at 48h control n⁼18, SkQ1 n⁼15, MitoTEMPO n⁼15; at 72h control n⁼18, SkQ1 n⁼14, MitoTEMPO n⁼15. Data points shown as mean +/- SEM. Supplementary Fig. 6. Assessment of Cytochrome C release in the liver of placebo-treated control vs. SkQ1-treated group at 24h post-CLP. CLP mice received total of three SkQ1/placebo injections before sacrifice at 24h. Data is shown as densitometric analysis of the Western blot from cytosolic. Total number of CLP mice loaded on three different gels: SkQ1 n⁼16; control (placebo) n⁼12. Data as (min-to-max) box-and-whiskers plots. Dotted lines indicate upper/lower standard deviation calculated based on eight healthy control mice (no CLP, no treatment) that were analyzed together with the CLP mice.CM:.coumassie stained gel.

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